This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is part of a Roadmap Consortium (U54 RR02437, T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women". The goal is to provide leadership in state-of-the-art cryopreservation technology to practitioners who are preserving human ovarian tissues under the aegis of the National Physicians Cooperative. The specific aim is to optimize ovarian cryopreservation methods in nonhuman primates for restoring fertility in female cancer survivors by comparing novel vitrification freeze-thaw methods to the currently utilized slow freeze method. Both slow freeze and vitrification of ovarian tissue in women have lead to pregnancies following autologous transplant. However, autologous transfer in many cancer patients is contraindicated because of the risk of re-seeding the patient with her own cancer. In this group of patients, encapsulated 3-dimensional (3D) culture of isolated secondary follicles post-thaw may be the best option for producing mature follicles and fertilizable oocytes for fertility preservation. Since slow freezing damages secondary follicles, a freezing/cooling method that could preserve secondary follicles for encapsulated 3D culture would bridge a technical gap for patients to utilize frozen ovarian tissue outside of transplantation. The success of ovarian tissue preservation will include assessment of a) histological and immunocytochemical characteristics of remaining follicles;b) secondary follicle survival, growth and functional characteristics using encapsulated 3D culture;and c) fertilization of the matured oocyte, early embryonic development, and ultimately production of live offspring. This project is facilitated by interactions with the Biomaterials Core at Northwestern University. Expected advances will be transferred to clinical efforts at the Fertility Centers.